Human Subcutaneous Derived Stromal Vascular Fraction Endothelial Cells Display Venous and Arterial Markers in Culture and 3D Capillaries.

BACKGROUND: The Stromal Vascular Fraction (SVF) derived from human subcutaneous fat has attracted pivotal interest in clinical applications for its regenerative and anti-inflammatory properties. A deeper characterisation of the endothelial cells within the SVF, across both traditional and tissue-engineered culture systems, is essential for advancing our understanding of endothelial cell biology and enhancing regenerative medicine therapies, including skin substitutes. METHODS: This study investigates endothelial cells from the SVF of human subcutaneous white adipose tissue in 2D culture and 3D bioengineered skin models to better define their specific subtypes. Immunofluorescence analysis was used to assess the SVF, with particular focus on endothelial cells, including their ability to form capillary-like networks within type I collagen hydrogels. RESULTS: Analysis of the SVF-derived cells showed PLVAP-positive blood endothelial cells but no lymphatic endothelial cells. The blood endothelial cells could be discriminated into NR2F2- and CD62E-positive venous endothelial cells and NRP1-expressing arterial endothelial cells. Within the 3D hydrogels, the blood endothelial cells formed venous and arterial capillaries. CONCLUSION: We characterised endothelial cells from human subcutaneous SVF, identifying venous and arterial blood endothelial cells while confirming the absence of lymphatic endothelial cells in vitro. These findings underline that subcutaneous adipose tissue is an attractive cell source due to its ease of isolation and abundance of endothelial cells for skin tissue engineering and regenerative medicine in general.